Fig. 1
From: Evaluation of the yeast surface display system for screening of functional nanobodies
Immunofluorescence labeling of yeast cells. Anti-hen egg-white lysozyme nanobody (LYS Nb) was displayed using five different plasmids. The yeast cells were stained using either anti-HA tag mouse monoclonal antibody or anti-FLAG tag mouse monoclonal antibody as the primary antibody and Alexa Fluor® 488 (AF488)-conjugated anti-mouse monoclonal antibody as the secondary antibody. The gene cassettes in Table I for each display system of LYS Nb are described above micrographs. LYS Nb: yeast cells with an LYS Nb-encoding plasmid. Control: yeast cells with a control plasmid that had no nanobody sequence and a FLAG-tag instead of an HA tag. Micrographs of a display system 1, b display system 2, c display system 3, d display system 4, and e display system 5. pGAP, glyceraldehyde-3-phosphate dehydrogenase promoter; pGAL1, galactokinase promoter; G. A. s. s., secretion signal of glucoamylase from Rhizopus oryzae; α-factor s. s., improved secretion signal of mating factor α preprotein from Saccharomyces serevisiae; Nb, anti-hen egg-white lysozyme nanobody; α-agglutinin, 320 C-terminal amino acids of α-agglutinin gene; 649-stalk, a synthetic anchor protein consisting of 649 amino acids. Scale bars, 10 μm